In many biological assays, especially those measuring fluorescence, instruments are restricted to a small number of possible simultaneous analysis channels, wherein only one analyte can be measured per channel. The existing methods of directly conjugating molecular detection complexes (i.e. antibodies) with different measurable labels (i.e. fluorescent dyes) require a significant investment of time and reagents. As a result, it is economically undesirable for manufacturers to produce and carry all possible combinations of directly-conjugated detection complexes. To circumvent this problem, in the field of flow cytometry unlabeled primary antibodies are commonly used in combination with directly labeled, isotype-specific secondary antibodies, but this approach is limited by the number of unique antibody isotypes in the reaction. Accordingly, there is a need for a multiplex-compatible labeling method that will allow manufacturers to minimize the number of unique reagents that must be synthesized, while conferring greater flexibility to the end user in terms of the number of different assays that can be designed using a standard set of reagents.
It is therefore one object of the disclosed invention to provide a method of rapidly and specifically conjugating molecular probes with detectable labels in an efficient protocol, via a convenient reaction, using stable reagents and materials, for the benefit of the end user's purposes.
It is another object of the disclosed invention to provide a method of rapidly and specifically conjugating multiple classes of molecular probes with different detectable labels in a multiplex reaction.
It is another object of the disclosed invention to allow the above to be carried out with reagents that, as individual components, are more stable and develop the desired novel characteristics (binding and detection), upon appropriate combination at a time more proximal to the experimentation.